Method for detecting mild impaired glucose tolerance or insulin hyposecretion

a glucose tolerance and insulin technology, applied in the field of mild impaired glucose tolerance or insulin secretion detection, can solve the problems of insufficient detection lower limit, difficult to detect, and difficult to achieve the effect of simple determination and good reproducibility

Active Publication Date: 2005-09-29
ASAHI KASEI PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0017] The present invention intends to provide an assay method for simple determination of

Problems solved by technology

Assays of blood glucose levels and insulin levels for those diagnoses are invasive procedures that require blood drawing more than once within a short time, giving the subjects considerable pains.
Nevertheless, the data are problematic in reproducibility and reliability because they varied among different examiners.
However, the myo-inositol assay reagent used in the report (c) has problems including: (i) an insufficient lower limit of detection because of a narrow measurement range and the need of diluting a sample for determination of various ur

Method used

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  • Method for detecting mild impaired glucose tolerance or insulin hyposecretion
  • Method for detecting mild impaired glucose tolerance or insulin hyposecretion
  • Method for detecting mild impaired glucose tolerance or insulin hyposecretion

Examples

Experimental program
Comparison scheme
Effect test

reference example 1

Study on thio-NAD Levels

[0157] 1) Reagents

[0158][0159] 5 mM MES (2-Morpholinoethanesulfonic acid) (pH 6.0) [0160] 0 to 40 mM thio-NAD (Oriental Yeast Co., Ltd.)

[0161][0162] 200 mM Bicine (pH 9.0) [0163] 0.3 mM NADH (Oriental Yeast Co., Ltd.) [0164] 25 u / mL myo-inositol dehydrogenase (Asahi Kasei Corporation)

[0165] 2) Method

[0166] The measurement device used was Autoanalyzer 7170S (Hitachi Chemical Co., Ltd.). To 3 μL of myo-inositol solution at concentrations of 0 to 3,000 μM, 180 μL of R-1 reagent was added and incubated at 37° C. for 4.8 minutes, followed by the addition of 180 μL of R-2 reagent to start the reaction. Absorbances at 405 nm were measured at 5.4 and 7.8 minutes after the reaction initiation, and then the difference therebetween was obtained. An increasing rate of absorbance per minute (ΔmABS / min) was calculated and the sensitivity was then investigated with respect to the standard solution.

[0167] 3) Results

[0168] The results are shown in FIG. 1. As shown in F...

reference example 2

Study on Buffers in Reagents for myo-Inositol Quantitative Determination

[0169] 1) Reagents

[0170][0171] 5 mM MES (pH 6.0) [0172] 5 mM thio-NAD (Oriental Yeast Co., Ltd.)

[0173][0174] 100 mM Buffer (pH 8.8) [0175] 0.5 mM NADH (Oriental Yeast Co., Ltd) [0176] 10 u / mL myo-inositol dehydrogenase (Asahi Kasei Corporation)

[0177] 2) Method

[0178] R-2 reagents for myo-inositol quantitative determination were prepared following the above shown composition with a buffer being selected from:

[0179] Tris (Tris(hydroxymethyl)aminomethane),

[0180] Tricine (N-Tris(hydroxymethyl)methylglycine),

[0181] Bicine (N,N-Bis(hydroxyethyl)glycine),

[0182] TAPS (N-Tris(hydroxymethyl)methyl-3-aminopropanesulfonic acid),

[0183] TEA (Triethanolamine),

[0184] CHES (2-Cyclohexylamino)ethanesulfonic acid), and

[0185] AMPSO(3-((1,1-Dimethyl-2-hydroxy-ethyl)amino)-2-hydroxypropanesulfonic acid).

[0186] The measurement device used was Autoanalyzer 7170S (Hitachi Chemical Co., Ltd.). To 15 μL of the standard 100-μM ...

reference example 3

Study on ADP-Hexokinase

[0189] 1) Reagents [0190][0191] 5 mM MES (pH 6.0) [0192] 5 mM MgCl2 (Wako Pure Chemical Industries, Ltd.) [0193] 8 mM ATP (Oriental Yeast Co., Ltd.) [0194] 10 mM thio-NAD (Oriental Yeast Co., Ltd.) [0195] 10 u / mL ATP-hexolinase (Asahi Kasei Corporation) [0196] 0 to 4 u / mL ADP-hexokinase (Asahi Kasei Corporation)

[0197][0198] 200 mM Bicine (pH 9.0) [0199] 0.3 mM NADH (Oriental Yeast Co., Ltd.) [0200] 25 u / mL myo-inositol dehydrogenase (Asahi Kasei Corporation)

[0201] 2) Method

[0202] The measurement device used was Autoanalyzer 7170S (Hitachi Chemical Co., Ltd.). Samples were prepared by mixing 100 μL of 2,000 μM myo-inositol solution and 1 mL of 0 to 10 g / dL glucose solution. To 3 μL of each sample, 180 μL of R-1 reagent was added and incubated at 37° C. for 4.8 minutes, followed by the addition of 180 μL of R-2 reagent to start the reaction. Absorbances at 405 nm were measured at 5.4 and 7.8 minutes after the reaction initiation, and then the difference ther...

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Abstract

It is intended to provide a noninvasive method of conveniently detecting mild impaired glucose tolerance and/or insulin hyposecretion at the early stage with the use of an enzyme. Namely, mild impaired glucose tolerance and/or hyposecretion at the early stage are detected by quantifying myoinositol secreted into the urine before loading glucose and after loading glucose for a definite period of time with the use of a reagent and comparing the increase (or the increase ratio) in the myoinositol content thus measured with a characteristic level which has been preliminarily determined in normal subjects.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a method of examining mild impaired glucose tolerance or insulin secretory defect using a sample such as urine. In addition, the present invention can be applied to a method for predicting or diagnosing a disease that stems from mild impaired glucose tolerance or insulin secretory defect, such as diabetes meritus, arteriosclerosis, or hypertension; a method of determining effects of prevention of, treatment of, or medical advice on those diseases; and a method of evaluating therapeutic agents for treatment of those diseases. BACKGROUND OF THE INVENTION [0002] A final goal of diabetic treatment is to prevent the onset of diabetic complications and to inhibit the development thereof As demonstrated by clinical tests for achieving this goal it is important to find any abnormality and start treatment thereof at the earliest possible stage [e.g., Diabetes Research and Clinical Practice, 28, 103 (1995)]. [0003] Further, it is ...

Claims

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Application Information

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IPC IPC(8): C12Q1/32C12Q1/48C12Q1/54G01N33/66G01N33/68
CPCC12Q1/32C12Q1/485G01N2800/042G01N33/66G01N33/6893C12Q1/54
Inventor YAMAKOSHI, MASARUKOUZUMA, TAKUJI
Owner ASAHI KASEI PHARMA
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